Enhanced MSC preparations

Patent metadata

Field	Value
Publication number	US-9963678-B2
Application number	US-201514662344-A
Country	US
Kind code	B2
Filing date	Mar 19, 2015
Priority date	Oct 8, 2010
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

The present invention provides preparations of MSCs with important therapeutic potential. The MSC cells are non-primary cells with an antigen profile comprising less than about 1.25% CD45+ cells (or less than about 0.75% CD45+), at least about 95% CD105+ cells, and at least about 95% CD166+ cells. Optionally, MSCs of the present preparations are isogenic and can be expanded ex vivo and cryopreserved and thawed, yet maintain a stable and uniform phenotype. Methods are taught here of expanding these MSCs to produce a clinical scale therapeutic preparations and medical uses thereof.

First claim

What is claimed is:

1. A mesenchymal stem cell (MSC) preparation comprising:
a. at least 1 billion cultured human bone marrow-derived MSCs expanded from a cryopreserved in-process intermediate MSC preparation; and
b. an antigen and activity profile comprising:
i. less than 0.75% CD45+ cells;
ii. at least 95% CD105+ cells;
iii. at least 95% CD166+ cells; and
iv. capable of inhibiting IL2Rα expression by CD3/CD28-activated PBMCs by at least 30% relative to a control.

2. The MSC preparation of claim 1 , wherein the at least 1 billion cultured human bone marrow-derived MSCs are isogenic.

3. The MSC preparation of claim 2 , wherein the MSC preparation comprises at least 20 billion cultured human bone marrow-derived MSCs in number.

4. The MSC preparation of claim 1 , wherein the MSCs express about 13 pg to about 44 pg TNFRI per million MSCs and wherein the MSCs inhibit IL2Rα expression by CD3/CD28-activated PBMCs when mixed with peripheral blood mononuclear cells (PBMCs) at a ratio of about 5 PBMCs per MSC.

5. The MSC preparation of claim 4 , wherein the MSCs are isogenic and are at least 4.5×10 9 human bone marrow-derived MSCs in number.

6. The MSC preparation of claim 5 , further comprising a cryopreservative.

7. The MSC preparation of claim 6 , wherein after a freeze-thaw cycle, at least 70% of the MCSs are viable, as assessed by dye exclusion.

8. The MSC preparation of claim 5 , wherein the MSCs:
a. are capable of at least 1 population doubling; and
b. retain said antigen and activity profile after the population doubling.

9. The MSC preparation of claim 5 , wherein the MSCs:
a. are capable of at least 1 population doubling; and
b. retain the differentiation capacity after the population doubling.

10. The MSC preparation of claim 1 , wherein the preparation contains less than 55 μg/ml BSA and less than 42 μg/ml trypsin.

11. The MSC preparation of claim 1 , wherein the preparation is substantially free of:
a. mycoplasma, endotoxin, and fungi; and
b. HTLV1, HTLV2, HBV, CMV, EBV, HHV-6A, HHV-6B, HHV-8, HIV, Parvovirus B-19, HCV, and HPV Type 18 viral nucleic acids.

Assignees

• Mesoblast Int Sarl
• Mesoblast Int Sarl

Inventors

• Tom Samson
• Ton Christopher
• Danilkovitch Alla

Classifications

• A61P5/14

  of the thyroid hormones, e.g. T3, T4 · CPC title

• A61P37/06

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• A61P35/00

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• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• A61P1/00

  Drugs for disorders of the alimentary tract or the digestive system · CPC title